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AB273747

Human CDH1 (E Cadherin) knockout A-431 cell line

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CDH1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein Frameshift = 95%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
5 Images
Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)
  • WB

Lab

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)

Lanes 1 - 4:

Western blot - Anti-E Cadherin antibody [SP64] (<a href='/en-us/products/primary-antibodies/e-cadherin-antibody-sp64-ab227639'>ab227639</a>) at 1/100 dilution

Lanes 1 - 4:

Western blot - Anti-E Cadherin antibody [SP64] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/e-cadherin-antibody-sp64-bsa-and-azide-free-ab240984'>ab240984</a>) at 1/100 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)

Lane 2:

CDH1 knockout A431 cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

MDA-MB-231 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 105 kDa,130 kDa

false

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)
  • WB

Lab

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)

Lanes 1 - 4:

Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (<a href='/en-us/products/primary-antibodies/e-cadherin-antibody-ep700y-intercellular-junction-marker-ab40772'>ab40772</a>) at 1/10000 dilution

Lanes 1 - 4:

Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (<a href='/en-us/products/primary-antibodies/e-cadherin-antibody-ep700y-low-endotoxin-azide-free-ab201499'>ab201499</a>) at 1/10000 dilution

Lanes 1 - 4:

Western blot - Anti-E Cadherin antibody [EP700Y] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/e-cadherin-antibody-ep700y-bsa-and-azide-free-ab256580'>ab256580</a>) at 1/10000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)

Lane 2:

CDH1 knockout Raji cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

MDA-MB-231 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 105 kDa,130 kDa

false

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)
  • WB

Lab

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)

Western blot : Anti-CDH1 antibody [4A2] (ab231303) staining at 1 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab231303 was shown to bind specifically to CDH1. A band was observed at 130, 110, 55 kDa in wild-type A431 cell lysates with no signal observed at this size in CDH1 knockout cell line ab273747 (knockout cell lysate ab273781). To generate this image, wild-type and CDH1 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-E Cadherin antibody [4A2] (<a href='/en-us/products/primary-antibodies/e-cadherin-antibody-4a2-ab231303'>ab231303</a>) at 1 µg/mL

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)

Lane 2:

CDH1 knockout A431 cell lysate at 20 µg

Lane 3:

Caco-2 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Predicted band size: 97 kDa

false

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)
  • WB

Lab

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)

Western blot : Anti-CDH1 antibody [EP700Y] (ab40772) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40772 was shown to bind specifically to CDH1. A band was observed at 130, 110, 80, 55, 40 kDa in wild-type A431 cell lysates with no signal observed at this size in CDH1 knockout cell line ab273747 (knockout cell lysate ab273781). To generate this image, wild-type and CDH1 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (<a href='/en-us/products/primary-antibodies/e-cadherin-antibody-ep700y-intercellular-junction-marker-ab40772'>ab40772</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)

Lane 2:

Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)

Lane 2:

CDH1 knockout A431 cell lysate at 20 µg

Lane 3:

Caco-2 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 97 kDa

false

Next Generation Sequencing - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)
  • NGS

Lab

Next Generation Sequencing - Human CDH1 (E Cadherin) knockout A-431 cell line (AB273747)

1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein Frameshift = 95%

Disease

Epidermoid Carcinoma

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Complementary Products

Cell Lines & Lysates

AB273834

Human SNX1 knockout HeLa cell line

Next Generation Sequencing - Human SNX1 knockout HeLa cell line (AB273834)

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Cell Lines & Lysates

AB276092

Human TP53 (p53) knockout A549 cell line

Advanced Validation

Western blot - Human TP53 (p53) knockout A549 cell line (AB276092)

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Primary Antibodies

AB231303

Anti-E Cadherin antibody [4A2]

BOND RX™ Validated|KO Validated|Lab Essentials

Western blot - Anti-E Cadherin antibody [4A2] (AB231303)

  • 4
  • 4
Assay Kits

AB233611

Human E-Cadherin ELISA Kit

Recombinant|SimpleStep

ELISA - Human E-Cadherin ELISA Kit (AB233611)

  • 5
  • 1
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Reactivity data

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Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
CDH1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

E-Cadherin sometimes called CDH1 or Cadherin-1 is a protein that plays a role in cell-to-cell adhesion. This transmembrane protein has a molecular weight of approximately 120 kDa. E-Cadherin is mainly expressed in epithelial tissues of various organs including the skin and gut. Its adhesive function is made possible by its extracellular domain that facilitates homophilic binding between cells contributing to the maintenance of tissue architecture and cellular integrity.
Biological function summary

E-Cadherin participates in establishing and maintaining adherens junctions which are vital for tissue structure. E-Cadherin operates as a core component of the cadherin-catenin complex which links the protein to the actin cytoskeleton. Through this linkage E-Cadherin plays an important role in signaling pathways that influence cellular growth and differentiation. The protein's ability to mediate intercellular connections also regulates cellular motility and supports basic aspects of cell behavior in epithelial tissues.

Pathways

E-Cadherin influences both the Wnt signaling pathway and the epithelial-to-mesenchymal transition (EMT). Within the Wnt signaling pathway E-Cadherin partners with β-catenin a significant player in transcription regulation and cell signaling. Disruption in E-Cadherin's adhesive functionality can lead to increased β-catenin availability affecting downstream transcriptional control. In the EMT process E-Cadherin loss characterizes an important step in which cells gain migratory and invasive properties typically seen during metastasis in cancer progression.

E-Cadherin's role is prominent in cancer particularly in the context of gastric and breast cancers. Mutations or altered expression of E-Cadherin lead to diminished cell adhesion promoting tumorigenesis and metastatic spread. In gastric cancer for instance mutated E-Cadherin often associates with loss of epithelial function facilitating cancer cell dissemination. Additionally its loss or dysfunction may correlate with proteins such as β-catenin further impacting cancer progression and pathology.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information CDH1
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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